PT  - JOURNAL ARTICLE
AU  - Martin Pabst
AU  - Denis Grouzdev
AU  - Christopher E. Lawson
AU  - Hugo B.C. Kleikamp
AU  - Carol de Ram
AU  - Rogier Louwen
AU  - Yuemei Lin
AU  - Sebastian Lücker
AU  - Mark C.M. van Loosdrecht
AU  - Michele Laureni
TI  - A general approach to explore prokaryotic protein glycosylation reveals the unique surface layer modulation of an anammox bacterium
AID  - 10.1101/2020.12.03.409086
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.12.03.409086
4099  - http://biorxiv.org/content/early/2020/12/03/2020.12.03.409086.short
4100  - http://biorxiv.org/content/early/2020/12/03/2020.12.03.409086.full
AB  - The enormous chemical diversity and strain variability of prokaryotic protein glycosylation makes a large-scale exploration exceptionally challenging. Therefore, despite the universal relevance of protein glycosylation across all domains of life, the understanding of their biological significance and the evolutionary forces shaping oligosaccharide structures remains highly limited.Here, we report on a newly established mass binning glycoproteomics approach that establishes the chemical identity of the carbohydrate components and performs untargeted exploration of prokaryotic oligosaccharides from large-scale proteomics data directly. We demonstrate our approach by exploring an enrichment culture of the globally relevant anaerobic ammonium-oxidizing bacterium Ca. Kuenenia stuttgartiensis. By doing so we resolved a remarkable array of oligosaccharides, produced by two entirely unrelated glycosylation machineries targeting the same surface-layer protein (SLP) simultaneously. More intriguingly, the investigated strain also accomplished modulation of highly specialized sugars, supposedly in response to its energy metabolism—the anaerobic oxidation of ammonium —which depends on the acquisition of substrates of opposite charge. Ultimately, we provide a systematic approach for the compositional exploration of prokaryotic protein glycosylation, and reveal for the first time a remarkable balance between maximising cellular protection through a complex array of oligosaccharides and adhering to the requirements of the ‘metabolic lifestyle’.Competing Interest StatementThe authors have declared no competing interest.